Daily rhythms in the hypothalamus-pituitary-interrenal axis and acute stress responses in a teleost flatfish, Solea senegalensis.

The endocrine axis controlling the stress response displays daily rhythms in many factors such as adrenal sensitivity and cortisol secretion. These rhythms have mostly been described in mammals, whereas they are poorly understood in teleost fish, so that their impact on fish welfare in aquaculture remains unexplored. In the present research, the authors investigated the daily rhythms in the hypothalamus-pituitary-interrenal (HPI) axis in the flatfish Solea senegalensis, which has both scientific and commercial interest. In a first experiment, hypothalamic expression of corticotropin-releasing hormone (crh) and its binding protein (crhbp), both pituitary proopiomelanocortin A and B (pomca and pomcb) expression, as well as plasma cortisol, glucose, and lactate levels were analyzed throughout a 24-h cycle. All variables displayed daily rhythms (cosinor, p < .05), with acrophases varying depending on the factor analyzed: crh and cortisol peaked at the beginning of the dark phase (zeitgeber time [ZT] = 14.5 and 14.4 h, respectively), pomca and pomcb as well as glucose at the beginning of the light phase (ZT = 1.2, 2.4, and 3.4 h, respectively), and crhbp and lactate at the end of the dark phase (ZT = 22.3 and 23.0 h, respectively). In a second experiment, the influence of an acute stressor (30 s of air exposure), applied at two different time points (ZT 1 and ZT 13), was tested. The stress response differed depending on the time of day, showing higher cortisol values (96.2 ± 10.7 ng/mL) when the stressor was applied at ZT 1 than at ZT 13 (52.6 ± 11.1 ng/mL). This research describes for the first time the daily rhythms in endocrine factors of the HPI axis of the flatfish S. senegalensis, and the influence of daytime on the stress responses. A better knowledge of the chronobiology of fish provides a helpful tool for understanding the circadian physiology of the stress response, and for designing timely sound protocols to improve fish welfare in aquaculture.

Metamorphosis induces a light-dependent switch in Senegalese sole (Solea senegalensis) from diurnal to nocturnal behavior.

Light plays a key role in the development of biological rhythms in fish. Recent research in Senegal sole has revealed that spawning and hatching rhythms, larval development, and growth performance are strongly influenced by lighting conditions. However, the effect of light on the daily patterns of behavior remains unexplored. Therefore, the aim of this study was to investigate the impact of different photoperiod regimes and white, blue, and red light on the activity rhythms and foraging behavior of Solea senegalensis larvae up to 40 days posthatching (DPH). To this end, eggs were collected immediately after spawning during the night and exposed to continuous white light (LL), continuous darkness (DD), or light-dark (LD) 12L:12D cycles of white (LD(W)), blue (LD(B), λ(peak) = 463 nm), or red light (LD(R), λ(peak) = 685 nm). A filming scenario was designed to video record activity rhythms during day and night times using infrared lights. The results revealed that activity rhythms in LD(B) and LD(W) changed from diurnal to nocturnal on days 9 to 10 DPH, coinciding with the onset of metamorphosis. In LD(R), sole larvae remained nocturnal throughout the experimental period, while under LL and DD, larvae failed to show any rhythm. In addition, larvae exposed to LD(B) and LD(W) had the highest prey capture success rate (LD(B) = 82.6% ± 2.0%; LD(W) = 75.1% ± 1.3%) and attack rate (LD(B) = 54.3% ± 1.9%; LD(W) = 46.9% ± 3.0%) during the light phase (ML) until 9 DPH. During metamorphosis, the attack and capture success rates in these light conditions were higher during the dark phase (MD), when they showed the same nocturnal behavioral pattern as under LD(R) conditions. These results revealed that the development of sole larvae is tightly controlled by light characteristics, underlining the importance of the natural underwater photoenvironment (LD cycles of blue wavelengths) for the normal onset of the rhythmic behavior of fish larvae during early ontogenesis.

Does lighting manipulation during incubation affect hatching rhythms and early development of sole?

Light plays a key role in the development of biological rhythms in fish. Previous research on Senegal sole has revealed that both spawning rhythms and larval development are strongly influenced by lighting conditions. However, hatching rhythms and the effect of light during incubation are as yet unexplored. Therefore, the aim of this study was to investigate the impact of the light spectrum and photoperiod on Solea senegalensis eggs and larvae until day 7 post hatching (dph). To this end, eggs were collected immediately after spawning during the night and exposed to continuous light (LL), continuous darkness (DD), or light-dark (LD) 12L:12D cycles of white light (LD(W)), blue light (LD(B); λ(peak) = 463 nm), or red light (LD(R); λ(peak) = 685 nm). Eggs exposed to LD(B) had the highest hatching rate (94.5% ± 1.9%), whereas LD(R) and DD showed the lowest hatching rate (54.4% ± 3.9% and 48.4% ± 4.2%, respectively). Under LD conditions, the hatching rhythm peaked by the end of the dark phase, but was advanced in LD(B) (zeitgeber time 8 [ZT8]; ZT0 representing the onset of darkness) in relation to LD(W) and LD(R) (ZT11). Under DD conditions, the same rhythm persisted, although with lower amplitude, whereas under LL the hatching rhythm split into two peaks (ZT8 and ZT13). From dph 4 onwards, larvae under LD(B) showed the best growth and quickest development (advanced eye pigmentation, mouth opening, and pectoral fins), whereas larvae under LD(R) and DD had the poorest performance. These results reveal that developmental rhythms at the egg stage are tightly controlled by light characteristics, underlining the importance of reproducing their natural underwater photoenvironment (LD cycles of blue wavelengths) during incubation and early larvae development of fish.

Synchronisation to light and feeding time of circadian rhythms of spawning and locomotor activity in zebrafish.

Reproduction in most fish is a seasonal phenomenon, since spawning occurs at a precise moment of the year to ensure maximal survival of the offspring. Nevertheless, fish reproduction cannot be considered an exclusively annual phenomenon, since spawning might also show daily rhythmicity. In this study, we used an automatic programmable egg collector to investigate the existence of circadian spawning and activity rhythms in zebrafish (Danio rerio L.), and their synchronization to different light and feeding cycles. Under 14L:10D, the results showed a diurnal spawning rhythm with an acrophase at ZT3 (lights went on at ZT0). Activity rhythms were also diurnal (74.4% of the total daily activity occurring during daytime), peaking immediately after lights on, in anticipation of spawning. Feeding at night did not change the diurnal spawning rhythm, but altered the daily pattern of activity, whose diurnal percentage dropped to 49.6%. When applying 1 h of darkness at ZT3, fish shifted the time of spawning to ZT7, while 1 h of darkness applied at ZT7 resumed spawning to ZT3. Under continuous light, locomotor activity rhythms persisted with tau=22.3 h and the spawning rhythm maintained its phase relationship with them, with an acrophase at CT3. In short, these findings revealed the existence of circadian spawning and locomotor rhythms in zebrafish. The two rhythms are in phase with each other and both are synchronized by light, though only locomotion is influenced by feeding time.

Effects of feeding schedule on locomotor activity rhythms and stress response in sea bream.

Feeding cycles entrain biological rhythms, which enable animals to anticipate feeding times and so maximize food utilization and welfare. In this article, the effect of mealtime was investigated in two groups of sea bream (Sparus aurata): one group received a single daily meal at random times during the light period (random daytime feeding, RDF), whereas the other group received the meal during the light period but at the same time (scheduled daytime feeding, SDF). All the fish showed diurnal behavior, although the SDF group showed a lower percentage of diurnalism (84.4% vs. 79.5% in RDF and SDF respectively) and developed food anticipatory activity some hours before the mealtime. In addition, the mean daily locomotor activity of the RDF group was significantly higher than that of the SDF group (3132 vs. 2654 counts/day, respectively). Although the mean weight differed between both groups on day 30 (115.7 g and 125.6 g in RDF and SDF respectively), these differences had disappeared by day 60. Plasma cortisol and glucose significantly differed in both groups (cortisol: 71.8 vs. 8.7 ng/ml, glucose: 53.7 vs. 43.8 mg/dl in RDF and SDF, respectively), whereas lactate did not differ significantly. The results obtained suggest that altering the feeding time (scheduled vs. random) affects the behavior and physiology of sea bream, indicating that a single daily feeding cycle (compared to random) is beneficial for fish welfare because they can prepare themselves for the forthcoming feed.